Last data update: 24 January 2024 16:39 CET
Plasmid name: phIL6h5HT1AR (LMBP 3027)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p3027.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human interleukin 6 genomic DNA (IL6, IL-6, IFNB2, BSF2, GeneID 3569) Human 5-hydroxytryptamine receptor 1A (HTR1A, 5-HT1A, GeneID 3350) |
| Promoter: | Human interleukin 6 promoter (IL6) Escherichia coli lac operon promoter |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Mammalian cells |
| Parental clone: | pBLHIL6CAT; pSP64H5HT1AR |
| Further information: | The plasmid was constructed by ligation of three fragments: the human 5HT1α receptor was isolated as a TfiI (filled in with Klenow DNA polymerase) / BamHI fragment from pSP64H5HT1AR. pBLHIL6CAT was XhoI opened, filled in with Klenow DNA polymerase and afterwards digested with BbsI. From this digestion the human interleukin 6 promoter (hIL6) and bacterial sequences were isolated. The third fragment contained the SV40 intron and SV40 polyadenylation sequences isolated from pBLHIL6CAT as a XhoII/BbsI fragment. When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide. The nucleotide sequence of the genomic hIL6 DNA corresponds with the EMBL Nucleotide Sequence Database accession number AC073072.11. However, the C nucleotide at promoter position -174 (polymorphism mutant) was changed into a G nucleotide (wild type). Name mentioned in Vanhoenacker et al. (1999) is pIL6-5HT1A. Other name of the plasmid is pHIL65HT1A. |
| EMBL Accession number: | AC073072.11, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 30/04/2004 |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed in the research unit of Em. Prof. Dr G. Haegeman(2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Vanhoenacker et al., Gene Ther. Mol. Biol. 3 (1999), 301-310 |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061 |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
phIL6h5HT1AR (LMBP 3027) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and was published in Vanhoenacker et al., 1999. |
Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.